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. 2021 Jun 25;12(7):649. doi: 10.1038/s41419-021-03920-4

Fig. 1. TNF-α induces endothelial–mesenchymal transition.

Fig. 1

A TNF-α decreases the protein expression of vascular endothelial marker (CD31) and increases the protein expression of mesenchymal markers (S100A4, α-SMA). HMVECs were treated for 96 h with 0, 20, 50 or 100 ng/ml of TNF-α. Protein expressions were analysed by western blot. B Morphological changes induced by a 100 ng/ml TNF-α treatment (photographs). The histogram shows ImageJ software analysis of cell morphology by calculating the elliptical form factor EFF (the major axis divided by minor axis). Results are the average of 100 cells. C TNF-α decreases the mRNA (left panel) and protein (right panel) expression of vascular endothelial markers (CD31, CD34, VE-cadherin) and increases the mRNA (left panel) and protein (right panel) expression of mesenchymal markers (COL1A1, N-cadherin, S100A4, α-SMA, SM22-α). mRNA levels were quantified by RT-qPCR. TBP for RT-qPCR and β-tubulin for western blot analysis were used as controls. RT-qPCR histograms show the mean of three independent biological experiments and western blots are representative of three independent biological experiments. Significant differences are indicated by solid lines (*P < 0.1, **P < 0.05, ***P < 0.005 by t test). Scale bars, 100 µm (inset) or 2000 µm. HMVEC human microvascular endothelial cell, TNF-α tumour necrosis factor-α, α-SMA α-smooth muscle actin, VE-cad VE-cadherin, COL1A1 collagen type I α1, N-Cad N-cadherin, Ctrl control, mRNA messenger RNA, TBP TATA-box binding protein.